MicroRNA‑28‑3p inhibits angiotensin‑converting enzyme 2 ectodomain shedding in 293T cells treated with the spike protein of severe acute respiratory syndrome coronavirus 2 by targeting A disintegrin and metalloproteinase 17

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus that causes coronavirus disease 2019. Angiotensin-converting enzyme 2 (ACE2) is the SARS-CoV binding site and is ubiquitously expressed in endothelial cells of several organs, with the highest levels in the cardiovascular system, kidney and lungs. A disintegrin and metalloproteinase 17 (ADAM17) is involved in ectodomain shedding of ACE2. In the present study, reverse-transcription-quantitative PCR, transfection, TUNNEL assay, dual-luciferase activity assay and western blotting were conducted to investigate the effects of microRNA (miR)-28-3p on ADAM17-dependent shedding of the ACE2 ectodomain following treatment with the spike protein (S-protein) of SARS-CoV-2. It was found that miR-28-3p was significantly downregulated in 293T cells treated with 100 ng/ml of S-protein for 24 h at 37˚C, which led to upregulation of ADAM17. In addition, the expression of ADAM17 and miR-28-3p were negatively correlated based on Pearson's correlation test in 293T cells treated with S-protein for 24 h. Overexpression of miR-28-3p and inhibition of ADAM17 regulated 293T cell viability, apoptosis and ACE2 ectodomain shedding. It was also demonstrated that ADAM17 was the target gene of miR-28-3p and that miR-28-3p negatively regulated ADAM17 expression. Notably, the inhibition of ADAM17 expression blocked the effects of miR-28-3p inhibitor on proliferation, apoptosis and ACE2 ectodomain shedding in 293T cells treated with S-protein.The findings of the present study suggested that miR-28-3p inhibits ADAM17-dependent ACE2 ectodomain shedding in 293T cells treated with the S-protein of SARS-CoV-2, which suggested the potential therapeutic role of miR-28-3p mimic in the prevention and treatment of patients with SARS-CoV-2.

miRNA expression in COVID-19

“… ( Yang et al, 2021 ) miR-28-3p In vitro 293 T cells Disintegrin and metalloproteinase 17 and ADAM17 miR-28-3p inhibits ADAM17-dependent ACE2 ectodomain shedding, making it a potential target in the prevention and management of COVID-19 patients. ( Xu and Li, 2021 ) miR-155 In vitro Vero E6, Calu-3, Caco-2 and H1299 Human epithelial cell line Calu-3 Induction of miR-155 and stimulation of the innate immune responses in SARS-CoV-2 is twice as high as in SARS-CoV. ( Wyler et al, 2021 ) miR-7-5p, miR-24-3p, miR-145-5p, miR-223-3p In vivo Young group elderly group healthy group diabetic group 141 S protein The mentioned miRNAs are decreased in the elderly and diabetic groups and can directly inhibit the expression of S protein and the replication of SARS-CoV-2 virus.…”

COVID-19: Mechanisms, risk factors, genetics, non-coding RNAs and neurologic impairments

“… MiRNA Study design Object Gene-targets Biological function Reference miR-98 Bioinformatics In vitro HMVEC-L HUVEC TMPRSS2 Modulates TMPRSS2 expression in the endothelial cells [ 115 ] miR-1207-5p Bioinformatics In vitro Human alveolar and bronchial epithelial cells CSF1 Enhances inflammatory responses in COVID-19 patients, and promoting EMT, which can contribute to pulmonary fibrosis, a possible sequela of COVID-19. [ 116 ] miR-28-3p In vitro 293 T cells Disintegrin and ADAM17 Exerts its function on both cell viability and cell apoptosis (Inhibit miR-28-3p expression inhibited cell viability and promoted cell apoptosis during S-protein treatment) [ 117 ] miR-31, miR-29, miR-126, and miR-17 Bioinformatics In vitro Human serum samples ZMYM5, COL5A3, and CAMSAP1 These miRNAs have been down-regulated and the levels of their mRNA targets have been enhanced with the increase of disease grade [ 118 ] miR-148a In vitro HEK-293T and CHME3 USP33, IRF9, TNFα, NF-κB, and IFN-β Activate human microglia [ 119 ] let7b‐5p In vitro Naso‐oropharyngeal swabs and ATCC ACE2 and DPP4 Participates in the mechanisms of acquiring the virulence of the virus and perform as a therapeutic target for COVID‐19 [ 120 ] miR-125b-5p and miR-155-5p In vitro Human serum samples …”

“…These data provide a molecular explanation for the resistance of monocytes to COVID-19 infection and support the notion that innate immunity mediated by intracellular miRNAs may play a significant role in protecting cells of the immune system from COVID-19 infection. A brief summary of the major miRNAs and their regulatory effects involving signal pathways of COVID-19 mechanism is shown in Table 1 [ [115] , [116] , [117] , [118] , [119] , [120] , [121] , [122] , [123] , [124] ].

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An update on angiotensin-converting enzyme 2 structure/functions, polymorphism, and duplicitous nature in the pathophysiology of coronavirus disease 2019: Implications for vascular and coagulation disease associated with severe acute respiratory syndrome coronavirus infection

“…Other miRNAs predicted to bind to ACE2 mRNA 3’-UTR, such as miR-9-5p and miR-218-5p, were found to be differentially expressed in different cell types ( Pierce et al, 2020 ). Moreover, the repression of the Xu and Li, 2021 ; Figure 5 ).An in silico studies aimed at predicting miRNAs that regulate ACE2-related networks with a possible impact on COVID-19 outcome, suggests that the top miRNAs regulating ACE2 networks are miR-27a-3p, miR-26b-5p, miR-10b-5p, miR-302c-5p, hsa-miR-587, hsa-miR-1305, hsa-miR-200b-3p, hsa-miR-124-3p, and hsa-miR-16-5p ( Wicik et al, 2020 ). sACE2 shed into systemic circulation maintains its ability to generate Ang-(1–7).…”